Mechanical stressing of integrin receptors induces enhanced tyrosine phosphorylation of cytoskeletally anchored proteins.
Physical forces play a fundamental role in the regulation of cell function in many tissues, but little is known about how cells are able to sense mechanical loads and realize signal transduction. Adhesion receptors like integrins are candidates for mechanotransducers. We used a magnetic drag force device to apply forces on integrin receptors in an osteoblastic cell line and studied the effect on tyrosine phosphorylation as a biochemical event in signal transduction. Mechanical stressing of both the beta1 and the alpha2 integrin subunit induced an enhanced tyrosine phosphorylation of proteins compared with integrin clustering. Application of cyclic forces with a frequency of 1 Hz was more effective than a continuous stress. Using Triton X-100 for cell extraction, we found that tyrosine-phosphorylated proteins became physically anchored to the cytoskeleton due to mechanical integrin loading. This cytoskeletal linkage was dependent on intracellular calcium. To see if mechanical integrin stressing induced further downstream signaling, we analyzed the activation of mitogen-activated protein (MAP) kinases and found an increased phosphorylation of MAP kinases due to mechanical stress. We conclude that integrins sense physical forces that control gene expression by activation of the MAP kinase pathway. The cytoskeleton may play a key role in the physical anchorage of activated signaling molecules, which enables the switch of physical forces to biochemical signaling events.
Schmidt
C
C
Department of Internal Medicine, University of Rostock, 18055 Rostock, Germany.
Pommerenke
H
H
Dürr
F
F
Nebe
B
B
Rychly
J
J
D015703:Antigens, CD / Q000502:physiology*; D002118:Calcium / Q000378:metabolism; D017871:Calcium-Calmodulin-Dependent Protein Kinases / Q000378:metabolism; D002614:Chelating Agents / Q000494:pharmacology; D015638:Cytochalasin D / Q000494:pharmacology; D003598:Cytoskeletal Proteins / Q000378:metabolism*; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D039421:Integrin alpha2; D019012:Integrin beta1 / Q000502:physiology; D016023:Integrins / Q000502:physiology*; D010766:Phosphorylation; D010812:Physical Stimulation; D011485:Protein Binding / Q000187:drug effects; D011956:Receptors, Cell Surface / Q000502:physiology*; D011990:Receptors, Transferrin / Q000378:metabolism; D015398:Signal Transduction; D014443:Tyrosine / Q000378:metabolism; D014732:Vibration
D015703:Antigens, CD; D002614:Chelating Agents; D003598:Cytoskeletal Proteins; D039421:Integrin alpha2; D019012:Integrin beta1; D016023:Integrins; D011956:Receptors, Cell Surface; D011990:Receptors, Transferrin; C070379:1,2-bis(2-aminophenoxy)ethane N,N,N',N'-tetraacetic acid acetoxymethyl ester; D015638:Cytochalasin D; D014443:Tyrosine; D004533:Egtazic Acid; D017871:Calcium-Calmodulin-Dependent Protein Kinases; D002118:Calcium